浙南地区HBV基因型分布特征及其与慢性乙型肝炎患者血清HBV DNA水平和HBeAg表达的关系

目的分析浙南地区慢性乙型肝炎患者HBV基因型分布特征,探讨不同基因型与患者血清中HBV DNA水平及HBeAg表达的关系。方法收集2011年6月-2012年3月161例就诊的HBV DNA阳性患者血清标本,采用型特异性引物PCR方法对HBV进行基因分型并用实时荧光定量PCR方法测量HBV DNA含量(拷贝/ml)和ELISA方法检测HBeAg;对HBV DNA含量进行对数转换使其符合正态分布后,运用χ2检验分析基因型与HBeAg阳性率的关系,t检验分析B、C基因型患者HBV DNA水平的差异。结果 161例患者中,B型41例,占25.5%;C型118例,占73.3%;BC混合型2例,占1.2%。C型患者HBV DNA水平和HBeAg阳性率分别为(5.84±1.40)log10拷贝/ml和64.4%,高于B型的(5.49±1.33)log10拷贝/ml和53.7%,但差异均无统计学意义(P0.05)。结论浙南地区HBV基因型以C型为主,B型次之。C基因型HBV DNA水平与HBeAg阳性率均高于B基因型,肝硬化的发生与C基因型HBV密切相关。     

浙江温州地区1020例乙型肝炎患者HBV基因分型研究

目的:了解浙江温州地区乙肝病毒基因型分布,初步评价微流芯片分析仪在乙肝病毒基因分型中的应用特点.方法:用HBV基因型特异引物经聚合酶链反应扩增温州地区1 020例慢性乙型肝炎患者血清HBV,其扩增产物应用微流芯片检测.结果:该地区1 020例患者中966例有基因分型结果,其中B型20.49%(198/966)、C型66.46%(642/966),B、C混合型4.87%(47/966),C、B混合型6.21%(60/966),D型1.97%(19/966).结论:浙江温州地区乙型肝炎病毒基因型分布具有北方流行特点,微流芯片在同类检测方法中具有灵敏、准确、快速优势.     

黄石地区慢性无症状乙型肝炎病毒携带者的基因型分布

目的：了解湖北黄石地区慢性无症状乙型肝炎病毒（HBV）携带者HBV基因型的分布，及其与病毒复制水平、HBeAg表达的相关性。方法：选择黄石地区168例慢性无症状HBV携带者作为研究对象，HBV基因型采用PCR微板核酸杂交．ELISA方法检测；血清HBVDNA复制水平采用荧光定量PCR检测；HBV—M采用ELISA法检测。结果：168例慢性无症状HBV携带者中HBVDNA阳性者为114例（阳性率为67．9％），其基因型分布为B、C、D型以及这3种基因型组成的混合型，而未发现A、E、F基因型。其中以B型、C型为主，所占比例为62．6％和36．8％，D型及混合型比例均为5．3％。C基因型患者中，HBV DNA呈现高水平复制（10^7～10^8 Copies／ml）的患者比例为26．2％（11／42），B基因型为13．3％（8／60）（P〈0．05）。C基因型患者血清抗-HBe阳性率（40．5％）显著高于B基因型（15．0％）（P〈0．01）。结论：黄石地区存在HBV的B、C、D基因型，以及由它们组成的混合基因型；B基因型为优势基因型；C基因型与HBV高复制水平，以及基因变异相关。     

秦皇岛市136例慢性乙型肝炎患者拉米夫定的疗效及其与HBV基因型的关系

目的:探讨秦皇岛市慢性乙型肝炎(CHB)患者拉米夫定治疗疗效与基因型的关系。方法:136例CHB患者口服拉米夫定,100mg/次,1次/d,疗程48周,用药前采用PCR方法测定乙型肝炎病毒(HBV)A～D基因型。结果:秦皇岛市CHB患者基因型以C型为主,占75.74%,其次为B基因型占16.91%,B/C混合型占7.35%,B基因型在拉米夫定抗病毒治疗48周时显示HBV DNA阴转率、HBeAg血清转换率、ALT复常率、治疗有效率4方面均高于C型及B/C基因型(P0.05),B基因型HBV感染者有较低的YMDD变异发生率。结论:拉米夫定抗病毒疗效与基因型有关,HBV基因型测定可作为预测拉米夫定抗病毒疗效的指标。     

乙肝病毒基因型与干扰素近期疗效探讨

目的：探讨HBV基因型与干扰素近期疗效的关系。方法：分析240例HBVDNA阳性的患者基因型，比较不同基因型患者干扰素近期疗效。结果：240例HBV感染者中HBV基因B型144例，占60．0％，C型90例，占37．5％，B、C混合型6例，占2．5％；干扰素的疗效以B型为高。结论：干扰素近期疗效以HBV基因型B型为好。     

乙型肝炎病毒基因型对乙型肝炎病毒变异的影响

目的探讨乙型肝炎病毒(HBV)基因型对病毒前核心区(前C区,nt1896)及基本核心启动子(BCP,nt1762/1764)变异的影响.方法416例血清HBsAg阳性、HBV DNA定量大于1.0×104拷贝/ml的患者,采用微流基因芯片检测HBV基因型、前C区及BCP变异.结果416例HBV感染者中406例有基因分型结果:B型20.9%、C型65.9%、BC混合型10.8%,10例患者未分出基因型.302例为HBeAg(-)且HBV DNA( )患者,其中248例(82.12%)有前C区或BCP变异,41.06%为前C区变异,31.12?P变异,2种同时变异为9.94%.B型患者前C区变异率为22.9%(20/87),与C型患者前C区变异率39.4%(108/274)及B、C混合型变异率40.0%相比均有显著差异(P＜0.01).在BCP变异及双变异,C型患者变异率均大于B型患者,但差异无统计学意义(P＞0.05).B、C混合型患者前C区及BCP变异率与C型相似.结论HBV基因型可影响病毒前C区及BCP变异,以C型为著.     

乙型肝炎病毒基因型与YMDD变异的关系

目的:了解乙型肝炎病毒(HBV)基因型与YMDD变异之间的关系.方法:多对型特异性引物PCR扩增法对238例经拉米夫定治疗的慢性乙型肝炎患者进行HBV基因分型,直接序列分析;采用基因芯片检测YMDD及前C/BCP区变异.结果:238例患者中,检测出B基因型190例(79.8%),C基因型41例(17.2%),BC混合型7例(3.0%);发生YMDD变异44例,变异率为18.5%,其中B基因型33例,变异率为17.4%,C基因型8例,变异率为19.5%,BC混合型3例,变异率为42.4%,C基因型YMDD变异的发生率与B基因型相比,差异无显著性意义(P＞0.05).44例YMDD变异者中,30例同时存在L528M变异,7例联合前C区(nt 1 896)变异,13例联合BCP区(nt 1 762/1 764)双重突变.结论:本地区慢性乙型肝炎患者中,优势基因型为B型和C型,经拉米夫定治疗后YMDD变异的发生率在B型和C型差异无显著性意义,同时伴有L528M及前C/BCP区多重变异.     

乙型肝炎病毒基因的型特异引物结合型特异核苷酸分析

目的 研究HBV感染者HBV基因型分布状况,并建立一套适合HBV病毒株分型的简便可靠的分析方法.方法 通过对GenBank中全部S区基因序列(1000余条)进行比对,筛选A～H 8个基因型的型特异核苷酸.采用型特异引物PCR法分型,对238例慢性乙型肝炎患者所感染病毒中未能分型的病毒株再进行S区基因巢式PCR扩增、测序筛选出其特异性核苷酸从而判定其基因型.结果 238株HBV均得到分型.其中B型HBV感染者159例(男120例、女39例);C型69例(男52例、女17例);B+C混合型6例(男4例、女2例),B+D混合型4例(男3例、女1例).B型、C型、B+C和B+D昆合型检出率分别为66.8%、28.9%、2.5%和1.6%.未检出A、E、F、G、H基因型.结论 HBV感染以B、C基因型为主,B基因型高于C基因型.少数患者为B+C或B+D混合型感染.B、C基因型分布与性别无关(χ2=0.794,P＞0.05).     

山西省HBV基因型分布特点和临床意义

目的:了解山西省常见的乙型肝炎病毒(HBV)基因型,探讨HBV各基因型与血清HBV-DNA水平以及疾病进展的临床意义.方法:随机收集山西省HBV DNA阳性病例680例进行基因分型,并结合临床资料进行统计学分析.结果:680例HBV感染者中,有65例(9.6％)未测出基因型.在可检出基因型的615例患者中,基因型B、C、B/C分别占8.9％、82.6％、8.5％.C型、B/C型的HBV DNA载量明显高于B型(P＜0.05);各基因型与肝病的严重程度间差异有显著性意义(P＜0.05).结论:山西省HBV基因型中以C型为优势,其次为B/C和B型.基因C型和B/C型在慢性重型肝炎中较为常见,基因C型更易发生肝硬化和肝癌.     

RtL164V,一个可能与拉米夫定耐药相关的突变

目的 进一步了解HBV P基因结构与拉米夫定的疗效关系.方法 一步法扩增HBVP基因后进行测序,对比拉米夫定治疗应答,无应答和突破患者治疗前后血清HBV P基因结构的变化.结果 在无应答和突破患者中,2例B基因型和1例C基因型转变为B、C混合型HBV感染;1例患者血清HBV发生B→C基因型转换.3组中部分患者治疗前血清中检出YMDD变异株HBV,所有8例治疗无应答和突破患者在治疗过程中均出现YMDD变异.所有无应答患者治疗前后和突破患者突破时的血清HBV均出现rtL164V变异,而在应答者中未见.突破和无应答患者逆转录酶(rt)保守区还出现rt191L、rtK168R、rtH2ML、rtS256C 4个氨基酸替代.结论 YMDD基因序列变异不是产生拉米夫定临床耐药的惟一原因,rtL164V变异可能是一个新的与拉米夫定耐药相关的突变.     

Geographic distribution, virologic and clinical characteristics of hepatitis B virus genotypes in China

The significance of hepatitis B virus (HBV) genotypes for the heterogeneity of chronic HBV infection and severity of liver disease is not well understood. The aim of this study was to determine the distribution and virologic characteristics of HBV genotypes in China and possible association with the diversity of liver disease. The study includes 1096 chronic HBV carriers from nine provinces in China. We collected clinical and laboratory data and analysed the HBV strains in sera by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and nucleotide sequencing techniques. The most common HBV genotypes were B (41%) and C (53%), while genotypes A and D were also found. A North-South divide was identified in genotype B and C distribution - genotype C was predominant in northern China, while genotype B was more prevalent in southern provinces. Patients with genotype B were younger than those with genotype C, and had a lower prevalence of HBeAg - 65%vs 72%, respectively (P = 0.03). However, the severity of liver disease did not differ significantly between patients infected with genotype B or C - neither when comparing liver function tests (1024 patients), nor hepatic inflammation and fibrosis (264 patients). Amongst 47 patients with genotype D (by PCR-RFLP), 37 (79%) were infected with a new subtype (designated Dc), having a recombination fragment from genotype C precore/core region. This is the first large-scale HBV genotype study from China and convincing documentation of the North-to-South gradient of genotypes C vs B in this country. HBV DNA recombination over the surface and precore/core genes increases the diversity of HBV strains and may have diagnostic and clinical implications.     

慢性乙型肝炎病毒基因型与BCP区变异的临床研究

为了研究乙型肝炎病毒 (HBV)基因型与C基因启动子 (BCP)基因变异的关系 ,对 6 9例慢性乙型肝炎患者分别用聚合酶链反应 (PCR)—限制性片段长度多态性 (RFLP)技术和PCR微板核酸分子杂交技术 ,进行基因分型及HBVBCP基因变异检测。在 6 9例慢性乙型肝炎患者中 ,各基因型发生的BCP区A176 2T/G176 4A双突变分别为C型 18例 (4 3 9% )B型 3例 (16 7% )D型 2例 (2 0 % )。C型的BCP双突变率明显高于B型 ,两者相比有显著性差异(P <0 0 5 )。故可以认为乙型肝炎病毒基因型与BCP区双突变存在有一定的相关性。推测A176 2T/G176 4A双突变可能是造成C型患者比B型存在更严重肝损害的原因之一     

New HBV subgenotype D9, a novel D/C recombinant,identified in patients with chronic HBeAg‐negative infection in Eastern India

Genome diversity is a hallmark of hepatitis B virus (HBV), which allowed its classification into 10 genotypes (A–J) and numerous subgenotypes. Among them, Genotype D is currently segregated into eight subgenotypes (D1–D8). Here, we report the identification and characterization of a novel subgenotype within genotype D of HBV from chronic hepatitis B e antigen (HBeAg)‐negative patients of Eastern India. Phylogenetic tree analysis based on complete genome sequences revealed that six of 39 HBV/D isolates formed a distinct cluster supported by high bootstrap value and had nucleotide divergence >4% relative to the known D subgenotypes (D1–D8), justifying their assignment into a new subgenotype (D9). By comparing the amino acid sequences of the four ORFs of HBV/D9 with D1–D8, 36 specific residues, including a unique one (E₁₁₂ in the core region), were identified that could be considered as a signature of D9. Further analysis by Simplot, BootScan and jpHMM demonstrated that D9 resulted from a discrete recombination with genotype C over the precore–core region. This type of recombination has not been described previously as all C/D recombinants reported so far possessed genotype C backbones with mosaic fragments derived from HBV/D. Interestingly, compared to other subgenotypes of HBV/D, D9 isolates had a higher frequency of mutations (A1762T and G1764A) in the basal core promoter region that had been implicated in the development of hepatocellular carcinoma. Further investigations are needed to determine the overall prevalence and clinical significance of these newly characterized D9 strains and to assess the impact of inter‐genotypic recombination on viral properties.     

基因芯片技术检测西藏拉萨地区的乙型肝炎病毒基因型

目的:研究西藏拉萨地区乙型肝炎病毒基因型的分布与特点.方法:采集92份西藏拉萨地区乙型肝炎患者的血清,参照GenBank中HBV DNA序列设计寡核苷酸探针并制备HBV基因分型芯片,利用套式PCR扩增HBV S基因部分片段,结合基因芯片、DNA测序和BioEdit软件进行基因分型检测,并对其与乙肝标志物、DNA含量、性别和民族之间的关系进行分析.结果:在92例血清标本中,套式PCR检测73例HBV DNA阳性可进行基因分型检测.其中B型13例(17.8%),C型18例(24.7%),D型39例(53.4%)和B/D混合基因型3例(4.1%).统计学分析3种基因型分布在不同乙肝标志物阳性、不同DNA含量和不同性别之间无差异,但与民族存在统计学差异(x~2=7.179,P<0.05).B型以汉族为主(9/13),而C、D型以藏族为主(12/18、28/39).将基因芯片分型的B、C、D型和B/D混合型进行DNA序列分析,表明两种分型方法的结果完全一致.结论:PCR结合基因芯片技术可用于HBV基因分型.西藏拉萨地区HBV基因型包括B、C、D和B/D混合型,其中以D型为主.     

Association of core promoter/precore mutations and viral load in e antigen-negative chronic hepatitis B patients

Apart from core promoter A1762T/G1764A and precore G1896A mutations, other hepatitis B virus (HBV) mutants are detected in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B (CHB). The aim of this study was to determine the effects of those mutants on clinical manifestation and viral loads of genotypes B and C HBV. Seventy-nine HBeAg-negative CHB patients with hepatitis flare were enrolled in this study and their HBV precore/core region were sequenced. Serial biochemical profiles and viral loads were assessed and compared. Fifty-three patients (67%) were infected by genotype B HBV and 26 (33%) were infected by genotype C HBV. The clinical manifestation and HBV viral loads were comparable between the two groups. However, genotype B was significantly associated with precore G1896A mutation (92.5%), and more mutations within nucleotide 1809-1817 were detected in patients infected by genotype B as compared with those infected by genotype C (18.9%vs 3.8%). Most of the cases had mutations at the -2, -3 or -5 position from the precore AUG initiation codon. Triple core promoter mutations T1753C/A1762T/G1764A [corrected] appeared to be linked to genotype C rather than genotype B HBV (19.2%vs 1.9%; P = 0.013). In multivariate analysis, the presence of either triple core promoter 1753/1762/1764 mutation or nucleotide 1809-1817 mutation was the only factor associated with lower HBV viral load (<70 Meq/mL) (odds ratio = 9.01; 95% CI 1.11-71.43; P = 0.04). In conclusion, minor HBV variants with mutations in the core promoter and precore region were detectable in genotypes B and C. Such HBV variants are genotype specific and related to viraemia levels.     

重叠HCV感染对HBV／C基因热点变异的影响

为探讨乙型肝炎病毒（ＨＢＶ）重叠丙型肝炎病毒（ＨＣＶ）感染时ＨＣＶ对ＨＢＶ复制和基因变异的影响，采用套式聚合酶链反应（ＰＣＲ）与限制性片段长度多态性（ＲＦＬＰ）相结合。对１９例ＨＢＶ感染重叠ＨＣＶ感染（Ａ组）和３１例单独ＨＢＶ感染（Ｂ组）的慢怀肝病患者分析前Ｃ区密码２８终止是（Ａ８３）和Ｃ密码９７异亮氨酸变为亮氨酸变异Ｌ９７）。结果显示Ａ组第一次ＰＣＲ阳性率（１６％）明显低于Ｂ组（６５％）（Ｐ，０     

Hepatitis B-associated vasculitis in Alaska Natives: viral genotype, clinical and serologic outcome.

BACKGROUND: The highest incidence of hepatitis B virus (HBV)-associated vasculitis in the world has been reported in Alaska Natives. We examined the incidence of HBV-associated vasculitis before and after mass HBV vaccine immunization and the association between HBV genotype and vasculitis in a population-based cohort study in Alaska natives chronically infected with HBV. METHODS: Genotyping was performed in vasculitis cases and 644 hepatitis B-positive controls without vasculitis using polymerase chain reaction and sequencing of the S gene. Occurrence of HBV vasculitis from 1974 to 2004 was calculated. HBV vasculitis patients and controls were also tested for basal core promoter and precore mutations. RESULTS: Fifteen cases of HBV-associated vasculitis were identified: 13 (86%) had genotype D and one each genotype A and F. Genotype D was more commonly found in patients with vasculitis than controls [odd ratio (OR)=5.9, confidence interval (95% CI) 1.2, 21.8; P<0.015). CONCLUSIONS: HBV-associated vasculitis was associated with genotype D.     

Prevalence of Hepatitis B Virus Genotype D in Precore Mutants Among Chronic Liver Disease Patients from New Delhi,India

Hepatitis B is one of the most important causes of chronic viral hepatitis world wide. Mutations in the precore region of the hepatitis B virus (HBV) genome are frequently found in hepatitis B envelope antigen-negative cases. Data from India on the HBV genotype-associated distribution of precore mutations are limited. Our objective in this study was to genotype and detect the precore mutant with a point mutation from G to A at nucleotide 1896 using ligase chain reaction (LCR) and direct sequencing. A total of 115 cases of chronic liver disease were screened. The cases were evaluated on the basis of history, clinical examination, liver function profile, and serological test for HBV infection, which includes HBsAg, anti HBcIgG, HBeAg using commercially available ELISA kits. The cases, which were HBeAg+, HBeAg-, and HBV DNA+, were subjected to LCR and confirmed by direct sequencing. Of 115 chronic liver disease cases, 50 (43.5%) cases were HBV DNA positive. All cases were subjected to LCR; 11 (22%) cases confirmed the presence of precore mutants, while the remaining 39 (78%) were classified as the wild form of the virus. HBV genotyping by direct sequencing revealed that genotype D was predominant in both wild and mutant forms of the virus. We conclude that the HBV genotype distribution was not significantly different between precore mutants and the wild form of the virus (P>0.05). North Indian patients with genotype D were more likely to have persistent HBV infection with precore mutants. HBV genotypes correlate with the clinical outcome of chronic HBV infection.